mouse anti cdkn1a p21  (Santa Cruz Biotechnology)

Journal: bioRxiv

Article Title: A feed-forward loop between niche adenosine and Gzmk⁺ CD8 T cells propagates systemic inflammaging

doi: 10.64898/2026.03.18.712515

Figure Lengend Snippet: (A-D) The analysis of male 19 months WT or Gzmk -/- mice (male). (A) Staining of SA-β-gal in liver and hippocampus. (B) H&E staining in liver, lung and kidney. (C) SA-β-gal staining for WAT from indicated mice. (D) Western blot analysis of the expression of p16, p21 and p53 in liver. (E) The bioluminescence imaging of indicated mice (15 months, female) by injecting luciferase substrates. (F-G) CD8 T cells (4 x10 6 ) from old or old Gzmk -/- mice were adoptively transferred into young p16 Ink4a -luciferase reporter mice, one month later, the luciferase activity was measured (F), and the expression of the p16 -driven luciferase reporter in indicated tissues were measured by RT-qPCR.

Article Snippet: Primary antibodies used in this study: Mouse anti-CDKN2A/p16 (Santa Cruz Biotechnology, cat# sc-1661, 1:1000) Mouse anti-CDKN1A p21 (Santa Cruz Biotechnology, cat# sc-6246, 1:1000) Mouse anti-p53 (Santa Cruz Biotechnology, cat# sc-98, 1:1000) Mouse anti-GAPDH (Absin, abs830030,1:10000) Rabbit anti-γH2A.X (Cell Signaling Technology, cat# 9718S, 1:1000)

Techniques: Staining, Western Blot, Expressing, Imaging, Luciferase, Activity Assay, Quantitative RT-PCR

Journal: bioRxiv

Article Title: A feed-forward loop between niche adenosine and Gzmk⁺ CD8 T cells propagates systemic inflammaging

doi: 10.64898/2026.03.18.712515

Figure Lengend Snippet: (A, B) Primary human bladder fibroblasts were isolated from surgical excision from bladder cancer patients and subjected to GZMK treatment. (A) HBFs were treated with 100nM GZMK for 5 days, and SA-β-gal staining were performed. (B) HBFs were treated with 100nM GZMK in medium without FBS for one day, IL-6 and TNFα in supernatant were measured by ELISA. (C, D) MEFs were isolated and subjected to GZMK treatment as same with . (E-H) BMDMs were subjected to Gzmk or indicated inhibitors treatment. (E) BMDMs were stimulated with Gzmk for 30 minutes and phosphorylated ERK and p38 were determined by flow cytometry. (F) BMDMs were cultured with Gzmk or combined with indicated inhibitors for three days, SA-β-gal staining were performed. BMDMs were cultured with Gzmk or combined with indicated inhibitors for one day, expression of P16 (G) and P21 (H) were analyzed by flow cytometry, IL-6 and TNFα (I) in supernatant were measured by ELISA.

Article Snippet: Primary antibodies used in this study: Mouse anti-CDKN2A/p16 (Santa Cruz Biotechnology, cat# sc-1661, 1:1000) Mouse anti-CDKN1A p21 (Santa Cruz Biotechnology, cat# sc-6246, 1:1000) Mouse anti-p53 (Santa Cruz Biotechnology, cat# sc-98, 1:1000) Mouse anti-GAPDH (Absin, abs830030,1:10000) Rabbit anti-γH2A.X (Cell Signaling Technology, cat# 9718S, 1:1000)

Techniques: Isolation, Staining, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Cell Culture, Expressing

Journal: bioRxiv

Article Title: A feed-forward loop between niche adenosine and Gzmk⁺ CD8 T cells propagates systemic inflammaging

doi: 10.64898/2026.03.18.712515

Figure Lengend Snippet: Old mice (19 months) were administrated with SCH or PPACK for one-month. The level of IL-6 and TNFα (A), ALT and AST (B) in plasma from indicated mice were determined by ELSIA. (C) Staining of SA-β-gal in liver. (D) H&E staining of lungs from indicated mice. (E) Staining of SA-β-gal in hippocampus from indicated mice. (F-G) Western blot analysis of P16, P21 and P53 at liver and brain from indicated mice.

Article Snippet: Primary antibodies used in this study: Mouse anti-CDKN2A/p16 (Santa Cruz Biotechnology, cat# sc-1661, 1:1000) Mouse anti-CDKN1A p21 (Santa Cruz Biotechnology, cat# sc-6246, 1:1000) Mouse anti-p53 (Santa Cruz Biotechnology, cat# sc-98, 1:1000) Mouse anti-GAPDH (Absin, abs830030,1:10000) Rabbit anti-γH2A.X (Cell Signaling Technology, cat# 9718S, 1:1000)

Techniques: Clinical Proteomics, Staining, Western Blot

Journal: Scientific reports

Article Title: Sodium Tanshinone IIA Sulfonate alleviates vascular senescence in diabetic mice by modulating the A20-NFκB-NLRP3 inflammasome-catalase pathway.

doi: 10.1038/s41598-024-68169-1

Figure Lengend Snippet: Figure 1. STS alleviates senescence in diabetic aortas and HG-treated ECs and VSMCs. (A–J) Representative images and the summarized data of CAT (400 × , scale bar: 20 μm), ROSgreen staining fluorescence (15 × , scale bar: 1000 μm), p21 immunofluorescence (400 × , scale bar: 20 μm), SA-β-gal staining (40 × , scale bar: 200 μm), Masson staining (400 × , scale bar: 20 μm) in aortas; (K,L) The summarized data of aortas responding to Ach and SNP; The ECs and VSMCs were treated with low glucose (5 mM, LG) or high glucose (30 mM, HG) for 72 h, with or without STS (100 μM) treatment. (M–P) Representative Western blot gels and the summarized data of show the protein expression of A20, CAT in ECs and VSMCs; (Q) and (U) Representative images of p21 IHC (400 × , scale bar: 20 μm), ROSgreen staining (400 × , scale bar: 20 μm) and SA-β-gal staining (400 × , scale bar: 40 μm) in ECs and VSMCs; (R–T) and (V–X) The summarized data of show p21 positive area, integrated fluorescent density of ROSgreenin and the area of SA-β-gal positive in ECs and VSMCs. *P < 0.05 vs. Control (Ctrl) or LG; #P < 0.05 vs. db/db or HG group (n = 6 in mice, n = 3 in cells). In the aorta results Ctrl represents littermate (wild-type, WT) of db/db mice with no treatment. In the cellular results, STS showed no significant impact on senescence-related markers in the LG group cells. In our animal pre-experiment, we found that STS had no significant effect on CAT and p21 levels in the aorta of Ctrl (WT) animals (Supplemental Fig. 3A–D). Therefore, in the animal results, we did not include the intervention of STS in the Ctrl group.

Article Snippet: After treatment, fixed the cells with pre-cooled anhydrous ethanol for 30 min, then block with 1% hydrogen peroxide solution for 10 min. Next, incubate with the primary antibody, mouse anti-Waf1/Cip1/CDKN1A p21 antibody (1:100, sc-6246, Santa Cruz, Shanghai, China), overnight at 4 °C.

Techniques: Staining, Fluorescence, Immunofluorescence, Western Blot, Expressing, Control

Journal: Scientific reports

Article Title: Sodium Tanshinone IIA Sulfonate alleviates vascular senescence in diabetic mice by modulating the A20-NFκB-NLRP3 inflammasome-catalase pathway.

doi: 10.1038/s41598-024-68169-1

Figure Lengend Snippet: Figure 3. STS alleviates vascular senescence by inhibiting the NLRP3 inflammasome under diabetic condition. (A–H) Representative images and the summarized data of CAT (400 × , scale bar: 20 μm), p21 immunofluorescence (400 × , scale bar: 20 μm), SA-β-gal staining (40 × , scale bar: 200 μm), Masson staining (400 × , scale bar: 20 μm) in aortas; (I,J) The summarized data of aortas responding to Ach and SNP; (K,L) and (M,N) Representative Western blot gels and the summarized data show the expression of CAT in ECs and VSMCs; (O–Q) and (R–T) Representative images and the summarized data show the ROGgreen and SA-β-gal staining in ECs and VSMCs. *P < 0.05 vs. Control (Ctrl) or LG; #P < 0.05 vs. db/db or HG group (n = 6 in mice, n = 3 in cells).

Article Snippet: After treatment, fixed the cells with pre-cooled anhydrous ethanol for 30 min, then block with 1% hydrogen peroxide solution for 10 min. Next, incubate with the primary antibody, mouse anti-Waf1/Cip1/CDKN1A p21 antibody (1:100, sc-6246, Santa Cruz, Shanghai, China), overnight at 4 °C.

Techniques: Immunofluorescence, Staining, Western Blot, Expressing, Control

Journal: Scientific reports

Article Title: Sodium Tanshinone IIA Sulfonate alleviates vascular senescence in diabetic mice by modulating the A20-NFκB-NLRP3 inflammasome-catalase pathway.

doi: 10.1038/s41598-024-68169-1

Figure Lengend Snippet: Figure 4. STS suppresses the expression of NLRP3 and senescence by inhibiting of NFκB pathway in HG-treated ECs and VSMCs. (A–D) Representative Western blot gels and summarized data show the phosphorylation of IκBα and NFκB, and the expression of NLRP3 in ECs; (E–H) Representative Western blot gels and summarized data show the phosphorylation of IκBα and NFκB, and the expression of NLRP3 in VSMCs; (I–K) Representative images and the summarized data show p21 immunofluorescence (400 × , scale bar: 20 μm) and SA-β-gal staining (400 × , scale bar: 20 μm) in ECs; (L–N) Representative images and the summarized data show p21 immunofluorescence (400 × , scale bar: 20 μm) and SA-β-gal staining (400 × , scale bar: 20 μm) in VSMCs. *P < 0.05 vs. LG; #P < 0.05 vs. HG treated group (n = 3).

Article Snippet: After treatment, fixed the cells with pre-cooled anhydrous ethanol for 30 min, then block with 1% hydrogen peroxide solution for 10 min. Next, incubate with the primary antibody, mouse anti-Waf1/Cip1/CDKN1A p21 antibody (1:100, sc-6246, Santa Cruz, Shanghai, China), overnight at 4 °C.

Techniques: Expressing, Western Blot, Phospho-proteomics, Immunofluorescence, Staining

Journal: Scientific reports

Article Title: Sodium Tanshinone IIA Sulfonate alleviates vascular senescence in diabetic mice by modulating the A20-NFκB-NLRP3 inflammasome-catalase pathway.

doi: 10.1038/s41598-024-68169-1

Figure Lengend Snippet: Figure 5. STS suppresses the activation of NLRP3 inflammasome and vascular senescence by activating A20 under diabetic condition. (A,B) and (C,D) Representative IHC images of NLRP3 (400 × , scale bar: 20 μm) and CAT (400 × , scale bar: 20 μm) and the summarized data of NLRP3 and CAT positive area percentage in aortas; (E,F) Representative immunofluorescence images of p21 (400 × , scale bar: 20 μm) and the summarized data of p21 integrated density in aortas; (G,H) Representative images of SA-β-gal staining (40 × , scale bar: 200 μm) and the summarized data of SA-β-gal area percentage in aortas; (I,J) Representative images of Masson staining (400 × , scale bar: 20 μm) and the summarized data of collagen area percentage in aortas; (K,L) The summarized data show aortas responding to Ach and SNP; (M–P) Representative Western blot gels and summarized data show the NLRP3 phosphorylation at serine 194 site, the production of NLRP3 dimer, the phosphorylation of p65, the expression of CAT in ECs and VSMCs; (Q,R) The summarized data show the active caspase-1 in ECs and VSMCs; (S–V) Representative images of SA-β-gal staining (400 × , scale bar: 20 μm) and summarized data of SA-β-gal positive area percentage in ECs and VSMCs. *P < 0.05 vs. Control (Ctrl); #P < 0.05 vs. db/db or HG treated group (n = 6 in mice, n = 3 in cells).

Article Snippet: After treatment, fixed the cells with pre-cooled anhydrous ethanol for 30 min, then block with 1% hydrogen peroxide solution for 10 min. Next, incubate with the primary antibody, mouse anti-Waf1/Cip1/CDKN1A p21 antibody (1:100, sc-6246, Santa Cruz, Shanghai, China), overnight at 4 °C.

Techniques: Activation Assay, Immunofluorescence, Staining, Western Blot, Phospho-proteomics, Expressing, Control